Design and Validation of a Microarray for Detection, Hemagglutinin Subtyping, and Pathotyping of Avian Influenza Viruses

Gall, Astrid; Hoffmann, Bernd; Harder, Timm; Grund, Christian; Höper, Dirk W.; Beer, Martin

doi:10.1128/jcm.01330-08

Cited by 56 publications

(43 citation statements)

References 37 publications

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“…1 Recent developments, like low-density microarray analysis or combined rolling-circle and PCR amplification using padlock primers, are powerful tools to further speed up and expand AIV diagnostic possibilities where fully equipped laboratories are available. 3,4,12 Given, however, that the majority of outbreaks of currently circulating HPAIV H5N1 occurs in regions where such laboratories are only infrequently encountered, the need is urgent to implement highly specific and sensitive rapid molecular tests for detection of AIV in less well equipped laboratories. Such assays might also be used directly in the field as so-called pen site tests.…”

mentioning

confidence: 99%

Evaluation of Two Commercial Loop-Mediated Isothermal Amplification Assays for Detection of Avian Influenza H5 and H7 Hemagglutinin Genes

Postel

Letzel

Frischmann³

et al. 2010

J VET Diagn Invest

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Abstract. Real-time reverse transcription loop-mediated isothermal amplification (real-time RT-LAMP) holds substantial potential as a highly sensitive, specific, and easy-to-perform molecular technique for pathogen detection in clinical samples. In the current study, the analytical and diagnostic performance of 2 commercial realtime RT-LAMP kits, Avian Flu H5 and Avian Flu H7, in detecting Avian influenza virus (AIV) infections were evaluated and compared with validated real-time reverse transcription polymerase chain reaction (RT-PCR) assays using RNA from reference virus isolates of subtypes H5 (n 5 24) and H7 (n 5 25) and of phylogenetically related subtypes (n 5 20). When real-time RT-LAMP was carried out according to the recommendations of the manufacturer, 3 out of 24 H5 isolates and 8 out of 25 H7 reference strains were not detected. Prolonging the amplification phase resulted in detection of all H5 isolates but also in false positive detection of 2 non-H5 isolates. Real-time RT-LAMP specific to H7 failed to detect 2 H7 isolates after prolonged amplification. According to the examination of RNA log dilutions, the sensitivity of the real-time RT-LAMP assays, for a number of historic but also recent strains, was considerably lower compared with subtype-specific real-time RT-PCR assays. Application of the real-time RT-LAMP assays for analysis of diagnostic samples from wild birds confirmed their lower sensitivity. Commercial real-time RT-LAMP as tested in this study with a broad range of AIV H5 and H7 strains of phylogenetically diverse yet recent origin, holds some promise for routine veterinary diagnostic purposes, although real-time RT-LAMP was markedly more vulnerable to a reduction of detection limits because of strainspecific sequence variation than subtype-specific real-time RT-PCR.

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confidence: 99%

Evaluation of Two Commercial Loop-Mediated Isothermal Amplification Assays for Detection of Avian Influenza H5 and H7 Hemagglutinin Genes

Postel

Letzel

Frischmann³

et al. 2010

J VET Diagn Invest

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“…Avian influenza viruses (AIVs) are subdivided into groups of high and low pathogenicity. The presence of multiple basic amino acids at the endoproteolytical cleavage site of the HA is a molecular indicator for high pathogenicity, at least for infections of gallinaceous poultry (4,5,6,7,8). Influenza viruses of HA subtypes H5, H7, and H9 are commonly identified in terrestrial gallinaceous poultry (9, 10).…”

mentioning

confidence: 99%

Alterations in Hemagglutinin Receptor-Binding Specificity Accompany the Emergence of Highly Pathogenic Avian Influenza Viruses

Heider

Mochalova

Harder

et al. 2015

J Virol

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Highly pathogenic avian influenza viruses (HPAIVs) of hemagglutinin H5 and H7 subtypes emerge after introduction of lowpathogenic avian influenza viruses (LPAIVs) from wild birds into poultry flocks, followed by subsequent circulation and evolution. The acquisition of multiple basic amino acids at the endoproteolytical cleavage site of the hemagglutinin (HA) is a molecular indicator for high pathogenicity, at least for infections of gallinaceous poultry. Apart from the well-studied significance of the multibasic HA cleavage site, there is only limited knowledge on other alterations in the HA and neuraminidase (NA) molecules associated with changes in tropism during the emergence of HPAIVs from LPAIVs. We hypothesized that changes in tropism may require alterations of the sialyloligosaccharide specificities of HA and NA. To test this hypothesis, we compared a number of LPAIVs and HPAIVs for their HA-mediated binding and NA-mediated desialylation of a set of synthetic receptor analogs, namely, ␣2-3-sialylated oligosaccharides. NA substrate specificity correlated with structural groups of NAs and did not correlate with pathogenic potential of the virus. In contrast, all HPAIVs differed from LPAIVs by a higher HA receptor-binding affinity toward the trisaccharides Neu5Ac␣2-3Gal␤1-4GlcNAc␤ (3=SLN) and Neu5Ac␣2-3Gal␤1-3GlcNAc␤ (SiaLe c ) and by the ability to discriminate between the nonfucosylated and fucosylated sialyloligosaccharides 3=SLN and Neu5Ac␣2-3Gal␤1-4(Fuc␣1-3)GlcNAc␤ (SiaLe x ), respectively. These results suggest that alteration of the receptor-binding specificity accompanies emergence of the HPAIVs from their low-pathogenic precursors. A vian influenza is a highly contagious infection with influenza A viruses, with a worldwide occurrence in aquatic wild bird populations which represent the major natural hosts of these viruses. Influenza A viruses are classified into subtypes according to their surface glycoproteins: 18 hemagglutinin (HA) and 11 neuraminidase (NA) antigenic subtypes have been identified (1, 2, 3). Avian influenza viruses (AIVs) are subdivided into groups of high and low pathogenicity. The presence of multiple basic amino acids at the endoproteolytical cleavage site of the HA is a molecular indicator for high pathogenicity, at least for infections of gallinaceous poultry (4,5,6,7,8). Influenza viruses of HA subtypes H5, H7, and H9 are commonly identified in terrestrial gallinaceous poultry (9, 10). Only low-pathogenic AIVs (LPAIVs) of subtypes H5 and H7 naturally evolve into highly pathogenic AIVs (HPAIVs), which cause severe infections with high rates of mortality in poultry (11,12,13,14) and can also be transmitted from birds to humans (15,16,17,18).The two viral glycoproteins, HA and NA, expressed on the surface of influenza virions play an important role in determining the pathogenic properties of the virus. Influenza virus infection is initiated by interactions between the viral HA and sialic acid-containing oligosaccharides on target cells. The NA cleaves off the terminal si...

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“…The antibodies against influenza surface antigens like HA and NA are used for the assays. PCR and RT-PCR based methods using specific primers against viral nucleic acids were more advanced, specific and comparatively faster methods of influenza virus detection but suffer from low throughput and limited multiplexing [6][7][8]. As influenza viruses have multiple types and subtypes which show high amount of genetic variation and mutability, sequencing or protein blotting based analysis is further needed to verify the PCR results.…”

Section: Microarray Chips For Identification and Sub Typing Of Influementioning

confidence: 99%

“…These microarrays use sets of immobilized oligonucleotide probes that form complexes with virus specific cDNA tagged with, a fluorescent label [8,11,12]. These microchips are used for the determination of different influenza virus types [13] and also for sub-typing majority of HA and NA influenza virus variants [21].…”

Section: Microarray Designing and Applicationmentioning

confidence: 99%

Impact of Microarray Technology in Influenza Virus Research and Diagnostics

Mukherjee¹

2013

J Proteomics Bioinform

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Design and Validation of a Microarray for Detection, Hemagglutinin Subtyping, and Pathotyping of Avian Influenza Viruses

Cited by 56 publications

References 37 publications

Evaluation of Two Commercial Loop-Mediated Isothermal Amplification Assays for Detection of Avian Influenza H5 and H7 Hemagglutinin Genes

Evaluation of Two Commercial Loop-Mediated Isothermal Amplification Assays for Detection of Avian Influenza H5 and H7 Hemagglutinin Genes

Alterations in Hemagglutinin Receptor-Binding Specificity Accompany the Emergence of Highly Pathogenic Avian Influenza Viruses

Impact of Microarray Technology in Influenza Virus Research and Diagnostics

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